Cat. # | Size | Qty. | Price |
---|---|---|---|
15956S | 100 µl (50 tests) |
|
REACTIVITY | H |
SENSITIVITY | Endogenous |
MW (kDa) | |
Source/Isotype | Rabbit IgG |
Product Information
Application | Dilution |
---|---|
Flow Cytometry (Fixed/Permeabilized) | 1:50 |
All reagents required for this protocol may be efficiently purchased together in our Intracellular Flow Cytometry Kit (Methanol) #13593, or individually using the catalog numbers listed below.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
NOTE: When including fluorescent cellular dyes in your experiment (including viability dyes, DNA dyes, etc.), please refer to the dye product page for the recommended protocol. Visit www.cellsignal.com for a full listing of cellular dyes validated for use in flow cytometry.
NOTE: Adherent cells or tissue should be dissociated and in single-cell suspension prior to fixation.
NOTE: Optimal centrifugation conditions will vary depending upon cell type and reagent volume. Generally, 150-300g for 1-5 minutes will be sufficient to pellet the cells.
NOTE: If using whole blood, lyse red blood cells and wash by centrifugation prior to fixation.
NOTE: Antibodies targeting CD markers or other extracellular proteins may be added prior to fixation if the epitope is disrupted by formaldehyde and/or methanol. The antibodies will remain bound to the target of interest during the fixation and permeabilization process. However, note that some fluorophores (including PE and APC) are damaged by methanol and thus should not be added prior to permeabilization. Conduct a small-scale experiment if you are unsure.
NOTE: Count cells using a hemocytometer or alternative method.
posted July 2009
revised June 2020
Protocol Id: 407
Human
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Glu320 of human COL11A1 protein.
The Extracellular Matrix (ECM) is a complex network of macromolecules that provides structural tissue support to cells in the basement membrane and interstitial matrix. It is composed of many molecules, including proteins, glycoproteins, proteoglycans, and polysaccharides (1,2). One of the major proteins that comprise the ECM, and the human body, is collagen. Collagens are a large family of proteins. They are trimeric molecules composed of three alpha polypeptide chains that form a triple helix structure that is characteristic of all collagens (3). The large family of collagens is divided into three subgroups: the fibrillar collagens, the non-fibril forming collagens, and the fibril-associated collagens. These subgroups differ in their structure and supramolecular assembly (3).
Collagen11A1 (COL11A1) is a minor fibrillar collagen that is not normally expressed at high levels in most normal tissues with the exception of cartilage, where it is expressed in high levels, and some other tissues/ organs, where it is expressed at a lower level (4). However, it has been reported that the expression of this molecule is correlated with advanced tumorigenic disease through meta analysis of data from multiple cancers, including ovarian, colon, breast, and lung (5). Additionally, it has also been associated with epithelial-mesenchymal transition (EMT) and metastasis (6,7). Cancer associated fibroblasts (CAFs) are typically the most abundant cell type in the stroma of many solid tumors. They are thought to contribute to ECM stiffness, which is ultimately thought to contribute to tumor growth and resistance to chemotherapeutic intervention. COL11A1 has been found to be elevated in CAFs and may contribute to chemotherapy resistance (8).
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